Creeping bentgrass Agrostis palustris (stolonifera) variety named &#39;T-1&#39;

ABSTRACT

A novel bentgrass cultivar, designated ‘T-1’, is disclosed. The invention relates to the seeds of bentgrass cultivar ‘T-1’, to the plants of bentgrass ‘T-1’ and to methods for producing a bentgrass plant produced by crossing the cultivar ‘T-1’ with itself or another bentgrass variety. The invention further relates to hybrid bentgrass seeds and plants produced by crossing the cultivar ‘T-1’ with another bentgrass cultivar.

BACKGROUND OF THE INVENTION

The present invention relates to a new and distinctive creepingbentgrass (Agrostis palustris (stolonifera)), a variety used primarilyas commercial golf course turf.

There are over 100 species of Bentgrass (Agrostis) but only two are usedto any great extent as golf course turf. Bentgrass is well adapted toclose mowing due to its prostrate growth habit. They grow best in moistuncompacted soils and have broad temperature hardiness.

The Agrostis genus—better known as the bentgrasses—is comprised of over100 species, several of which have been developed into successfulturfgrasses. One Agrostis in particular, A. stolonifera or creepingbentgrass, has become the preeminent grass for golf course puttinggreens the world over. Another Agrostis species, colonial bentgrass (A.tenuisSibth.), has been bred into a golf course grass useful on tees andfairways in cooler regions. Two or three other Agrostis species findminor turf application, mostly for golf, tennis courts, bowling greens,or an occasional home lawn.

The Agrostis genus is widely distributed throughout the world withrepresentative species found on all of the northern continents. However,of the present-day bentgrass species in use as turfgrasses, alloriginated from Europe. The original seed of these plants was brought tothe US during colonial times.

America has an abundance of native bentgrass species (A. S. Hitchcock,1951, Manual of the grasses of the United States. USDA Misc. Publ. 200)but none are commercially useable as turf grass.

Creeping bentgrass (Agrostis palustris) is so named due to its abilityto creep laterally by stolons. The stolons are able to root at the nodesproducing a new plant. Creeping bentgrass is the plant of choice forfairways, tees and greens where the height of cut is below one-halfinch.

Creeping bentgrass (Agrostis palustris) is a perennial cool season grassthat forms a dense mat. The grass spreads by profuse creeping stolonsand basal tillers and possesses rather vigorous, shallow roots. Stems,or stolons, are decumbent (creeping) and slender and produce long narrowleaves. Leaf blades are smooth on the upper surface and ridged on theunderside, are approximately 1 to 3 mm wide and bluish green inappearance. The ligule is long, membranous, finely toothed or entire androunded, auricles are absent.

Developing new grass species is difficult, time consuming, andexpensive. The developer must sift through thousands of prospectivegrasses listed in botanical literature, identify promising grasses, andoften travel thousands of miles to locate, isolate, identify, transport,quarantine, grow, test, and breed these grasses. This process can takemore than 10 years to develop acceptable cultivars. Furthermore, as itturns out, most prospective grasses in nature have no commercial turfvalue, due to their inability to generate an acceptable ground coverwhen mowed. The vast majority of natural grasses cannot produce a plushlawn under continuing defoliation.

Yet another complexity facing the plant developer is theunresponsiveness of many wild grasses to plant breeding. The vastmajority of wildland grasses lack genetic potential for refinement intodesirable turfgrass cultivars. Only after considerable investment incollection and breeding does the developer discover which grass speciescan be successful bred and which cannot.

The development of new turf grasses requires the development andselection of bentgrass varieties, the crossing of these varieties andselection of superior hybrid crosses. The hybrid seed is produced bymanual crosses between selected male-fertile parents or by using malesterility systems. These hybrids are selected for certain single genetraits such as pod color, flower color, pubescence color or herbicideresistance which indicate that the seed is truly a hybrid. Additionaldata on parental lines, as well as the phenotype of the hybrid,influence the breeder's decision whether to continue with the specifichybrid cross.

Pedigree breeding and recurrent selection breeding methods are used todevelop cultivars from breeding populations. Breeding programs combinedesirable traits from two or more cultivars or various broad-basedsources into breeding pools from which cultivars are developed byselfing and selection of desired phenotypes. The new cultivars areevaluated to determine which have commercial potential.

Pedigree breeding is used commonly for the improvement ofself-pollinating crops. Two parents that possess favorable,complementary traits are crossed to produce an F₁. An F₂ population isproduced by selfing one or several F₁'s. Selection of the bestindividuals may begin in the F₂ population; then, beginning in the F₃,the best individuals in the best families are selected. Replicatedtesting of families can begin in the F₄ generation to improve theeffectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new cultivars.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror inbred line which is the recurrent parent. The source of the trait tobe transferred is called the donor parent. The resulting plant isexpected to have the attributes of the recurrent parent (e.g., cultivar)and the desirable trait transferred from the donor parent. After theinitial cross, individuals possessing the phenotype of the donor parentare selected and repeatedly crossed (backcrossed) to the recurrentparent. The resulting plant is expected to have the attributes of therecurrent parent (e.g., cultivar) and the desirable trait transferredfrom the donor parent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F₂ plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

In a multiple-seed procedure, bentgrass breeders commonly harvest one ormore panicles from each plant in a population and thresh them togetherto form a bulk. Part of the bulk is used to plant the next generationand part is put in reserve. The procedure has been referred to asmodified single-seed descent or the panicle-bulk technique.

The multiple-seed procedure has been used to save labor at harvest. Itis considerably faster to thresh panicles with a machine than to removeone seed from each by hand for the single-seed procedure. Themultiple-seed procedure also makes it possible to plant the same numberof seeds of a population each generation of inbreeding. Enough seeds areharvested to make up for those plants that did not germinate or produceseed.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., Allard, 1960; Simmonds, 1979; Sneep et al., 1979; Fehr,1987).

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current cultivars. In addition toshowing superior performance, there must be a demand for a new cultivarthat is compatible with industry standards or which creates a newmarket. The introduction of a new cultivar will incur additional coststo the seed producer, the grower, processor and consumer; for specialadvertising and marketing, altered seed and commercial productionpractices, and new product utilization. The testing preceding release ofa new cultivar should take into consideration research and developmentcosts as well as technical superiority of the final cultivar. Forseed-propagated cultivars, it must be feasible to produce seed easilyand economically.

SUMMARY OF THE INVENTION

According to the invention, there is provided a novel cultivar ofbentgrass species for turf purposes. Cultivars from this speciesdemonstrate tolerance to close mowing, resistance to insects and fungaldiseases, high shoot density, ground coverage, resistance to Poa annua,wear tolerance and genetically dark green color.

Definitions

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Allele. Allele is any of one or more alternative forms of a gene, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing. “Backcrossing” is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotypes of the F₁hybrid.

Culm Length. “Culm Length” is defined as the length of the reproductivestem, measured from the crown growing point (below ground) to the tip ofthe inflorescence.

Liqule Length. “Ligule Length” is defined as the length of themembranous tissue protrusion on the adaxial side of the leaf collar atthe junction of the length blade and leaf sheath.

Flag Leaf Length and Width. “Flag leaf length and width” are thedimensions of the first leaf below the seed head (inflorescence).

Panicle Length. The term “panicle length” means the distance from thelowest branch to the tip of the inflorescence.

Plants headed. The term “plants headed” refers to the percent of plantsthat were at or past the boot stage of reproductive development on Jun.11, 2002 (an indicator of reproductive maturity).

Boot stage. The term “boot stage” refers to the stage of reproductionwhere the inflorescence emerges from the leaf sheath.

Plants in pollination. “Plants in pollination” refers to the percent ofplants in active pollination on Jun. 27, 2002 (another indicator ofreproductive maturity).

Plants with purple heads. “Plants with purple heads” refers to thepercent of plants with purple pigment in the seedhead (versus tan or nopurple) on Jun. 27, 2002.

DETAILED DESCRIPTION OF THE INVENTION

‘T-1’ is an improved, creeping bentgrass.

Some of the criteria used to select in various generations includedesirable dark green color, minimal thatch production, favorableresponse to mowing and competitive against Poa annua, seed yield,emergence, disease tolerance, maturity and plant height.

The cultivar has shown uniformity and stability, as described in thefollowing variety description information.

Bentgrass ‘T-1’ has the following morphologic and other characteristics(based primarily on data collected at Post Falls, Id.). TABLE 1 VARIETYDESCRIPTION INFORMATION Culm length (cm): 34.7 Flagleaf length (cm):3.50 Flagleaf width (mm): 2.47 Flagleaf length/width ratio (mm): 14.4Ligule length (mm): 1.94 Panicle length (cm): 6.67 Seed length (mm):1.58 Seed width (mm): 0.44 Seed length/width ratio (mm): 3.62

This invention is also directed to methods for producing a bentgrassplant by crossing a first parent bentgrass plant with a second parentbentgrass plant, wherein the first or second bentgrass plant is thebentgrass plant from the line ‘T-1’. Further, both first and secondparent bentgrass plants may be from the cultivar ‘T-1’. Therefore, anymethods using the cultivar ‘T-1’ are part of this invention: selfing,backcrosses, hybrid breeding, and crosses to populations. Any plantsproduced using cultivar ‘T-1’ as a parent are within the scope of thisinvention.

Useful methods include but are not limited to expression vectorsintroduced into plant tissues using a direct gene transfer method suchas microprojectile-mediated delivery, DNA injection, electroporation andthe like. More preferably, expression vectors are introduced into planttissues using the microprojectile media delivery with the biolisticdevice Agrobacterium-medicated transformation. Transformant plantsobtained with the protoplasm of the invention are intended to be withinthe scope of this invention.

The cultivar ‘T-1’ is similar to L-93. While similar to L-93, there arenumerous differences including: ‘T-1’ has a shorter mature plant heightand stature, higher shoot density under close mowing and darker greengenetic color.

FURTHER EMBODIMENTS OF THE INVENTION

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign genes, or additional, or modified versions of native, orendogenous, genes (perhaps driven by different promoters) in order toalter the traits of a plant in a specific manner. Such foreignadditional and/or modified genes are referred to herein collectively as“transgenes”. Over the last fifteen to twenty years several methods forproducing transgenic plants have been developed, and the presentinvention, in particular embodiments, also relates to transformedversions of the claimed variety or line.

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of or operatively linked to a regulatoryelement (for example, a promoter). The expression vector may contain oneor more such operably linked gene/regulatory element combinations. Thevector(s) may be in the form of a plasmid, and can be used alone or incombination with other plasmids, to provide transformed bentgrassplants, using transformation methods as described below to incorporatetransgenes into the genetic material of the bentgrass plant(s).

Expression Vectors for Bentgrass Transformation: Marker Genes—Expressionvectors include at least one genetic marker, operably linked to aregulatory element (a promoter, for example) that allows transformedcells containing the marker to be either recovered by negativeselection, i.e., inhibiting growth of cells that do not contain theselectable marker gene, or by positive selection, i.e., screening forthe product encoded by the genetic marker. Many commonly used selectablemarker genes for plant transformation are well known in thetransformation arts, and include, for example, genes that code forenzymes that metabolically detoxify a selective chemical agent which maybe an antibiotic or a herbicide, or genes that encode an altered targetwhich is insensitive to the inhibitor. A few positive selection methodsare also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) under the control of plantregulatory signals confers resistance to kanamycin. Fraley et al., Proc.Natl. Acad. Sci. U.S.A., 80:4803 (1983). Another commonly usedselectable marker gene is the hygromycin phosphotransferase gene whichconfers resistance to the antibiotic hygromycin. Vanden Elzen et al.,Plant Mol. Biol., 5:299 (1985).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, aminoglycoside-3′-adenyl transferase,the bleomycin resistance determinant. Hayford et al., Plant Physiol.86:1216 (1988), Jones et al., Mol. Gen. Genet., 210:86 (1987), Svab etal., Plant Mol. Biol. 14:197 (1990) Hille et al., Plant Mol. Biol. 7:171(1986). Other selectable marker genes confer resistance to herbicidessuch as glyphosate, glufosinate or broxynil. Comai et al., Nature317:741-744 (1985), Gordon-Kamm et al., Plant Cell 2:603-618 (1990) andStalker et al., Science 242:419423 (1988).

Other selectable marker genes for plant transformation are not ofbacterial origin. These genes include, for example, mouse dihydrofolatereductase, plant t-enolpyruvylshikimate-3-phosphate synthase and plantacetolactate synthase. Eichholtz et al., Somatic Cell Mol. Genet. 13:67(1987), Shah et al., Science 233:478 (1986), Charest et al., Plant CellRep. 8:643 (1990).

Another class of marker genes for plant transformation require screeningof presumptively transformed plant cells rather than direct geneticselection of transformed cells for resistance to a toxic substance suchas an antibiotic. These genes are particularly useful to quantify orvisualize the spatial pattern of expression of a gene in specifictissues and are frequently referred to as reporter genes because theycan be fused to a gene or gene regulatory sequence for the investigationof gene expression. Commonly used genes for screening presumptivelytransformed cells include α-glucuronidase (GUS, β-galactosidase,luciferase and chloramphenicol, acetyltransferase. Jefferson, R. A.,Plant Mol. Biol. Rep. 5:387 (1987), Teeri et al., EMBO J. 8:343 (1989),Koncz et al., Proc. Natl. Acad. Sci. USA 84:131 (1987), DeBlock et al.,EMBO J. 3:1681 (1984).

Recently, in vivo methods for visualizing GUS activity that do notrequire destruction of plant tissue have been made available. MolecularProbes publication 2908, Imagene Green™, p. 14 (1993) and Naleway etal., J. Cell Biol. 115:151a (1991). However, these in vivo methods forvisualizing GUS activity have not proven useful for recovery oftransformed cells because of low sensitivity, high fluorescentbackgrounds and limitations associated with the use of luciferase genesas selectable markers.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells. Chalfie et al., Science 263:802 (1994). GFP and mutants of GFPmay be used as screenable markers.

Promoters—Genes included in expression vectors must be driven bynucleotide sequence comprising a regulatory element, for example, apromoter. Several types of promoters are now well known in thetransformation arts, as are other regulatory elements that can be usedalone or in combination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred”.Promoters that initiate transcription only in certain tissue arereferred to as “tissue-specific”. A “cell-type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may effect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter that is active under mostenvironmental conditions.

Inducible Promoters—An inducible promoter is operably linked to a genefor expression in bentgrass. Optionally, the inducible promoter isoperably linked to a nucleotide sequence encoding a signal sequencewhich is operably linked to a gene for expression in bentgrass. With aninducible promoter the rate of transcription increases in response to aninducing agent.

Any inducible promoter can be used in the instant invention. See Ward etal., Plant Mol. Biol. 22:361-366 (1993). Exemplary inducible promotersinclude, but are not limited to, that from the ACEI system whichresponds to copper (Mett et al., PNAS 90:45674571 (1993)); In2 gene frommaize which responds to benzenesulfonamide herbicide safeners (Hersheyet al., Mol. Gen Genetics 227:229-237 (1991) and Gatz et al., Mol. Gen.Genetics 243:32-38 (1994) or Tet repressor from Tn10 (Gatz et al., Mol.Gen. Genetics 227:229-237 (1991). A particularly preferred induciblepromoter is a promoter that responds to an inducing agent to whichplants do not normally respond. An exemplary inducible promoter is theinducible promoter from a steroid hormone gene, the transcriptionalactivity of which is induced by a glucocorticosteroid hormone. Schena etal., Proc. Natl. Acad. Sci. USA 88:0421 (1991).

Constitutive Promoters—A constitutive promoter is operably linked to agene for expression in bentgrass or the constitutive promoter isoperably linked to a nucleotide sequence encoding a signal sequencewhich is operably linked to a gene for expression in bentgrass.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell et al., Nature 313:810-812 (1985) and the promoters from suchgenes as rice actin (McElroy et al., Plant Cell 2: 163-171 (1990));ubiquitin (Christensen et al., Plant Mol. Biol. 12:619-632 (1989) andChristensen et al., Plant Mol. Biol. 18:675-689 (1992)); PEMU (Last etal., Theor. Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J.3:2723-2730 (1984)) and maize H3 histone (Lepetit et al., Mol. Gen.Genetics 231:276-285 (1992) and Atanassova et al., Plant Journal 2 (3):291-300 (1992)).

The ALS promoter, Xba1/NcoI fragment 5′ to the Brassica napus ALS3structural gene (or a nucleotide sequence similarity to said Xba1/NcoIfragment), represents a particularly useful constitutive promoter. SeePCT application WO96/30530.

Tissue-specific or Tissue-preferred Promoters—A tissue-specific promoteris operably linked to a gene for expression in bentgrass. Optionally,the tissue-specific promoter is operably linked to a nucleotide sequenceencoding a signal sequence which is operably linked to a gene forexpression in bentgrass. Plants transformed with a gene of interestoperably linked to a tissue-specific promoter produce the proteinproduct of the transgene exclusively, or preferentially, in a specifictissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferredpromoter—such as that from the phaseolin gene (Murai et al., Science23:476-482 (1983) and Sengupta-Gopalan et al., Proc. Natl. Acad. Sci.USA 82:3320-3324 (1985)); a leaf-specific and light-induced promotersuch as that from cab or rubisco (Simpson et al., EMBO J.4(11):2723-2729 (1985) and Timko et al., Nature 318:579-582 (1985)); ananther-specific promoter such as that from LAT52 (Twell et al., Mol.Gen. Genetics 217:240-245 (1989)); a pollen-specific promoter such asthat from Zm13 (Guerrero et al., Mol. Gen. Genetics 244:161-168 (1993))or a microspore-preferred promoter such as that from apg (Twell et al.,Sex. Plant Reprod. 6:217-224 (1993).

Signal Sequences for Targeting Proteins to SubcellularCompartments—Transport of protein produced by transgenes to asubcellular compartment such as the chloroplast, vacuole, peroxisome,glyoxysome, cell wall or mitochondrion or for secretion into theapoplast, is accomplished by means of operably linking the nucleotidesequence encoding a signal sequence to the 5′ and/or 3′ region of a geneencoding the protein of interest. Targeting sequences at the 5′ and/or3′ end of the structural gene may determine, during protein synthesisand processing, where the encoded protein is ultimatelycompartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample, Becker et al., Plant Mol. Biol. 20:49 (1992), Close, P. S.,Master's Thesis, Iowa State University (1993), Knox, C., et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley”, Plant Mol. Biol. 9:3-17 (1987), Lerner et al., Plant Physiol.91:124-129 (1989), Frontes et al., Plant Cell 3:483-496 (1991), Matsuokaet al., Proc. Natl. Acad. Sci. 88:834 (1991), Gould et al., J. Cell.Biol. 108:1657 (1989), Creissen et al., Plant J. 2:129 (1991), Kalderon,et al., A short amino acid sequence able to specify nuclear location,Cell 39:499-509 (1984), Steifel, et al., Expression of a maize cell wallhydroxyproline-rich glycoprotein gene in early leaf and root vasculardifferentiation, Plant Cell 2:785-793 (1990).

Foreign Protein Genes and Agronomic Genes—With transgenic plantsaccording to the present invention, a foreign protein can be produced incommercial quantities. Thus, techniques for the selection andpropagation of transformed plants, which are well understood in the art,yield a plurality of transgenic plants which are harvested in aconventional manner, and a foreign protein then can be extracted from atissue of interest or from total biomass. Protein extraction from plantbiomass can be accomplished by known methods which are discussed, forexample, by Heney and Orr, Anal. Biochem. 114:92-6 (1981).

According to a preferred embodiment, the transgenic plant provided forcommercial production of foreign protein is a bentgrass plant. Inanother preferred embodiment, the biomass of interest is seed. For therelatively small number of transgenic plants that show higher levels ofexpression, a genetic map can be generated, primarily via conventionalRFLP, PCR and SSR analysis, which identifies the approximate chromosomallocation of the integrated DNA molecule. For exemplary methodologies inthis regard, see Glick and Thompson, Methods in Plant Molecular Biologyand Biotechnology CRC Press, Boca Raton 269:284 *1993). Map informationconcerning chromosomal location is useful for proprietary protection ofa subject transgenic plant. If unauthorized propagation is undertakenand crosses made with other germplasm, the map of the integration regioncan be compared to similar maps for suspect plants, to determine if thelatter have a common parentage with the subject plant. Map comparisonswould involve hybridizations, RFLP, PCR, SSR and sequencing, all ofwhich are conventional techniques.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary genes implicated in this regard include, but are notlimited to, those categorized below:

1. Genes That Confer Resistance to Pests or Disease and That Encode:

A. Plant disease resistance genes. Plant defenses are often activated byspecific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example Jones et al., Science 266:789 (1994)(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin et al., Science 262:1432 (1993) (tomato Pto gene for resistanceto Pseudomonas syringae pv. Tomato encodes a protein kinase); Mindrinoset al. Cell 78:1089 (1994) (Arabidopsis RSP2 gene for resistance toPseudomonas syringae).

B. A gene conferring resistance to a pest, such as soybean cystnematode. See e.g., PCT Application WO96/30517; PCT ApplicationWO93/19181.

C. A Bacillus thuringiensis protein, a derivative thereof or a syntheticpolypeptide modeled thereon. See, for example, Geiser et al., Gene48:109 (1986), who disclose the cloning and nucleotide sequence of a Btσ-endotoxin gene. Moreover, DNA molecules encoding σ-endotoxin genes canbe purchased from American Type Culture Collection, Manassas, Va., forexample, under ATCC Accession Nos. 40098, 67136, 31995 and 31998.

D. A lectin. See, for example, the disclose by Van Damme et al., PlantMolec. Biol. 24:25 (1994), who disclose the nucleotide sequences ofseveral Clivia miniata mannose-binding lectin genes.

E. A vitamin-binding protein such as avidin. See PCT applicationUS93/06487, the contents of which are hereby incorporated by reference.The application teaches the use of avidin and avidin homologues aslarvicides against insect pests.

F. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. See, for example, Abe et al., J. Biol. Chem.262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor), Huub et al., Plant Molec. Biol. 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor I), Sumitani etal., Biosci. Biotech. Biochem. 57:1243 (1993) (nucleotide sequence ofStreptomyces nitrosporeus α-amylase inhibitor) and U.S. Pat. No.5,494,813 (Hepher and Atkinson, issued Feb. 27, 1996).

G. An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

H. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosures of Regan, J. Biol. Chem. 269:9 (1994) (expression cloningyields DNA coding for insect diuretic hormone receptor), and Pratt etal., Biochem. Biophys. Res. Comm. 163:1243 (1989) (an allostatin isidentified in Diploptera puntata). See also U.S. Pat. No. 5,266,317 toTomalski et al., who disclose genes encoding insect-specific, paralyticneurotoxins.

I. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, see Pang et al., Gene 116:165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

J. An enzyme responsible for a hyperaccumulation of a monterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

K. An enzyme involved in the modification, including theost-translational modification, of a biologically active molecule; forexample, a glycolytic enzymes, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23:691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hookworm chitinase, and Kawalleck et al.,Plant Molec. Biol. 21:673 (1993), who provide the nucleotide sequence ofthe parsley ubi4-2 polyubiquitin gene.

L. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess etal., Plant Physiol. 104:1467. (1994), who provide the nucleotidesequence of a maize calmodulin cDNA clone.

M. A hydrophobic moment peptide. See PCT application WO95/16776(disclosure of peptide derivatives of Tachyplesin which inhibit fungalplant pathogens) and PCT application WO95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance), the respectivecontents of which are hereby incorporated by reference.

N. A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure of Jaynes et al., Plant Sci 89:43 (1993), ofheterologous expression of a cecropin-β lytic peptide analog to rendertransgenic tobacco plants resistant to Pseudomonas solanacearum.

O. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. rev. Phytopathol.28:451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virusand tobacco mosaic virus.

P. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor et al., Abstract #497, Seventh Int'l Symposium on MolecularPlant-Microbe Interactions (Edinburgh, Scotland) (1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

Q. A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366:469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

R. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-α-1,4-D-galacturonase. See Lamb et al., Bio/Technology10:1436 (1992). The cloning and characterization of a gene which encodesa bean endopolygalacturonase-inhibiting protein is described by Toubartet al., Plant J. 2:367 (1992).

S. A development-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10:305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

2. Genes That Confer Resistance to a Herbicide, For Example:

A. A herbicide that inhibits the growing point or meristem, such as animidazalinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as descried, for example, by Lee et al.,EMBO J. 7:1241 (1988), and Miki et al., Theor. Appl. Genet. 80:449(1990), respectively.

B. Glyphosate (resistance impaired by mutant5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase, PAT and Streptomyces hygroscopicusphosphinothricin-acetyl transferase, bar, genes), and pyridinoxy orphenoxy proprionic acids and cycloshexones (ACCase inhibitor-encodinggenes). See, for example, U.S. Pat. No. 4,940,835 to Shah, et al., whichdiscloses the nucleotide sequence of a form of EPSP which can conferglyphosate resistance. A DNA molecule encoding a mutant aroA gene can beobtained under ATCC accession number 39256, and the nucleotide sequenceof the mutant gene is disclosed in U.S. Pat. No. 4,769,061 to Comai.European patent application No. 0 333 033 to Kumada et al., and U.S.Pat. No. 4,975,374 to Goodman et al., disclose nucleotide sequences ofglutamine synthetase genes which confer resistance to herbicides such asL-phosphinothricin. The nucleotide sequence of aphosphinothricin-acetyl-transferase gene is provided in Europeanapplication No. 0 242 246 to Leemans et al., DeGreef et al.,Bio/Technology 7:61 (1989) describe the production of transgenic plantsthat express chimeric bar genes coding for phosphinothricin acetyltransferase activity. Exemplary of genes conferring resistance tophenoxy proprionic acids and cycloshexones, such as sethoxydim andhaloxyfop are the Acc1-S1, Acc1-S2, and Acc2-S3 genes described byMarshall et al., Theor. Appl. Genet. 83:435 (1992).

C. A herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Przibila et al.,Plant Cell 3:169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441 and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al., Biochem. J.285:173 (1992).

3. Genes That Confer or Contribute to a Value-Added Trait, Such as:

A. Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of stearoyle-ACP desaturase to increase stearicacid content of the plant. See Knultzon et al., Proc. Natl. Acad. Sci.USA 89:2625 (1992).

B. Decreased phytate content—1) Introduction of a phytase-encoding genewould enhance breakdown of phytate, adding more free phosphate to thetransformed plant. For example, see Van Hartingsveldt et al., Gene127:87 (1993), for a disclosure of the nucleotide sequence of anAspergillus niger phytase gene. 2) A gene could be introduced thatreduced phytate content. In maize, this, for example, could beaccomplished, by cloning and then reintroducing DNA associated with thesingle allele which is responsible for maize mutants characterized bylow levels of phytic acid. See Raboy et al., Maydica 35:383 (1990).

C. Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch. See Shiroza et al., J. Bacteriol. 170:810(1988) (nucleotide sequence of Streptococcus mutantsfructosyltransferase gene), Steinmetz et al., Mol. Gen. Genet. 20:220(1985) (nucleotide sequence of Bacillus subtilis levansucrase gene), Penet al., Bio/Technology 10:292 (1992) (production of transgenic plantsthat express Bacillus licheniformis α-amylase), Elliot et al., PlantMolec. Biol. 21:515 (1993) (nucleotide sequences of tomato invertasegenes), S{acute over (ø)}gaard et al., J. Biol. Chem. 268:22480 (1993)(site-directed mutagenesis of barley α-amylase gene), and Fisher et al.,Plant Physiol. 102:1045 (1993) (maize endosperm starch branching enzymeII).

Methods for Creeping Bentgrass Transformation—Numerous methods for planttransformation have been developed, including biological and physical,plant transformation protocols. See, for example, Miki et al.,“Procedures for Introducing Foreign DNA into Plants” in Methods in PlantMolecular Biology and Biotechnology, Glick, B. R. and Thompson, J. E.Eds. (CRC Press, Inc. Boca Raton, 1993) pages 67-88. In addition,expression vectors and in-vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available. See, forexample, Gruber et al., “Vectors for Plant Transformation” in Methods inPlant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J.E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages 89-119.

A. Agrobacterium-mediated Transformation—One method for introducing anexpression vector into plants is based on the natural transformationsystem of Agrobacterium. See, for example, Horsch et al., Science227:1229 (1985). A. tumefaciens and A. rhizogenes are plant pathogenicsoil bacteria which genetically transform plant cells. The Ti and Riplasmids of A. tumefaciens and A. rhizogenes, respectively, carry genesresponsible for genetic transformation of the plant. See, for example,Kado, C. I., Crit. Rev. Plant Sci. 10:1 (1991). Descriptions ofAgrobacterium vector systems and methods for Agrobacterium-mediated genetransfer are provided by Gruber et al., supra, Miki et al., supra andMoloney et al., Plant Cell Reports 8:238 (1989). See also, U.S. Pat. No.5,563,055 (Townsend and Thomas), issued Oct. 8, 1996.

B. Direct Gene Transfer—Several methods of plant transformationcollectively referred to as direct gene transfer, have been developed asan alternative to Agrobacterium-mediated transformation. A generallyapplicable method of plant surface of microprojectiles measuring 1 to 4μm. The expression vector is introduced into plant tissues with abiolistic device that accelerates the microprojectiles to speeds of 300to 600 m/s which is sufficient to penetrate plant cell walls andmembranes. Sanford et al., Part. Sci. Technol. 5:27 (1987), Sanford, J.C., Trends Biotech. 6:299 (1988), Klein et al., Bio/Tech. 6:559-563(1988), Sanford, J. C. Physiol Plant 7:206 (1990), Klein et al.,Biotechnology 10:268 (1992). See also U.S. Pat. No. 5,015,580 (Christou,et al.), issued May 14, 1991; U.S. Pat. No. 5,322,783 (Tomes, et al.),issued Jun. 21, 1994.

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang et al., Bio/Technology 9:996 (1991). Alternatively,liposome or spheroplast fusion have been used to introduce expressionvectors into plants. Deshayes et al., EMBO J., 4:2731 (1985), Christouet al., Proc Natl. Acad. Sci. USA 84:3962 (1987). Direct uptake of DNAinto protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-ornithine have also been reported. Hain et al., Mol. Gen. Genet.199:161 (1985) and Draper et al., Plant Cell Physiol. 23:451 (1982).Electroporation of protoplasts and whole cells and tissues have alsobeen described. Donn et al., In Abstracts of VIIth InternationalCongress on Plant Cell and Tissue Culture IAPTC, A2-38, p 53 (1990);D'Halluin et al., Plant Cell 4:1495-1505 (1992) and Spencer et al.,Plant Mol. Biol. 24:51-61 (1994).

Following transformation of creeping bentgrass target tissues,expression of the above-described selectable marker genes allows forpreferential selection of transformed cells, tissues and/or plants,using regeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic variety. The transgenic variety could then becrossed, with another (non-transformed or transformed) variety, in orderto produce a new transgenic variety. Alternatively, a genetic trait thathas been engineered into a particular soybean line using the foregoingtransformation techniques could be moved into another line usingtraditional backcrossing techniques that are well known in the plantbreeding arts. For example, a backcrossing approach could be used tomove an engineered trait from a public, non-elite variety into an elitevariety, or from a variety containing a foreign gene in its genome intoa variety or varieties that do not contain that gene. As used herein,“crossing” can refer to a simple X by Y cross, or the process ofbackcrossing, depending on the context.

Tissue Culture of Bentgrasss—When the term bentgrass plant is used inthe context of the present invention, this also includes any single geneconversions of that variety. The term single gene converted plant asused herein refers to those bentgrass plants which are developed by aplant breeding technique called backcrossing wherein essentially all ofthe desired morphological and physiological characteristics of a varietyare recovered in addition to the single gene transferred into thevariety via the backcrossing technique. Backcrossing methods can be usedwith the present invention to improve or introduce a characteristic intothe variety. The term backcrossing as used herein refers to the repeatedcrossing of a hybrid progeny back to the recurrent parent, i.e.,backcrossing 1, 2, 3, 4, 5, 6, 7, 8 or more times to the recurrentparent. The parental bentgrass plant that contributes the gene for thedesired characteristic is termed the nonrecurrent or donor parent. Thisterminology refers to the fact that the nonrecurrent parent is used onetime in the backcross protocol and therefore does not recur. Theparental bentgrass plant to which the gene or genes from thenonrecurrent parent are transferred is known as the recurrent parent asit is used for several rounds in the backcrossing protocol (Poehlman &Sleper, 1994; Fehr, 1987). In a typical backcross protocol, the originalvariety of interest (recurrent parent) is crossed to a second variety(nonrecurrent parent) that carries the single gene of interest to betransferred. The resulting progeny from this cross are then crossedagain to the recurrent parent and the process is repeated until abentgrass plant is obtained wherein essentially all of the desiredmorphological and physiological characteristics of the recurrent parentare recovered in the converted plant, in addition to the singletransferred gene from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single gene of the recurrent variety ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphological,constitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross, one ofthe major purposes is to add some commercially desirable, agronomicallyimportant trait to the plant. The exact backcrossing protocol willdepend on the characteristic or trait being altered to determine anappropriate testing protocol. Although backcrossing methods aresimplified when the characteristic being transferred is a dominantallele, a recessive allele may also be transferred. In this instance itmay be necessary to introduce a test of the progeny to determine if thedesired characteristic has been successfully transferred.

Many single gene traits have been identified that are not regularlyselected for in the development of a new variety but that can beimproved by backcrossing techniques. Single gene traits may or may notbe transgenic, examples of these traits include but are not limited to,male sterility, waxy starch, herbicide resistance, resistance forbacterial, fungal, or viral disease, insect resistance, male fertility,enhanced nutritional quality, industrial usage, yield stability andyield enhancement. These genes are generally inherited through thenucleus. Several of these single gene traits are described in U.S. Pat.Nos. 5,959,185; 5,973,234 and 5,977,445; the disclosures of which arespecifically hereby incorporated by reference.

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of bentgrass andregeneration of plants therefrom is well known and widely published. Forexample, reference may be had to Komatsuda, T. et al., “Genotype XSucrose Interactions for Somatic Embryogenesis in Soybean,” Crop Sci.31:333-337 (1991); Stephens, P. A., et al., “Agronomic Evaluation ofTissue-Culture-Derived Soybean Plants,” Theor. Appl. Genet. (1991)82:633-635; Komatsuda, T. et al., “Maturation and Germination of SomaticEmbryos as Affected by Sucrose and Plant Growth Regulators in SoybeansGlycine gracilis Skvortz and Glycine max (L.) Merr.” Plant Cell, Tissueand Organ Culture, 28:103-113 (1992); Dhir, S. et al., “Regeneration ofFertile Plants from Protoplasts of Soybean (Glycine max L. Merr.);Genotypic Differences in Culture Response,” Plant Cell Reports (1992)11:285-289; Pandey, P. et al., “Plant Regeneration from Leaf andHypocotyl Explants of Glycine-wightii (W. and A.) VERDC. var.longicauda,” Japan J. Breed. 42:1-5 (1992); and Shetty, K., et al.,“Stimulation of In Vitro Shoot Organogenesis in Glycine max (Merrill.)by Allantoin and Amides,” Plant Science 81:245-251 (1992); as well asU.S. Pat. No. 5,024,944 issued Jun. 18, 1991 to Collins et al., and U.S.Pat. No. 5,008,200 issued Apr. 16, 1991 to Ranch et al., the disclosuresof which are hereby incorporated herein in their entirety by reference.Thus, another aspect of this invention is to provide cells which upongrowth and differentiation produce creeping bentgrass plants having thephysiological and morphological characteristics of creeping bentgrassvariety ‘T-1’.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, plant clumps, and plantcells that can generate tissue culture that are intact in plants orparts of plants, such as embryos, pollen, flowers, seeds, pods, leaves,stems, roots, root tips, anthers, and the like. Means for preparing andmaintaining plant tissue culture are well known in the art. By way ofexample, a tissue culture comprising organs has been used to produceregenerated plants. U.S. Pat. Nos. 5,959,185; 5,973,234 and 5,977,445describe certain techniques, the disclosures of which are incorporatedherein by reference.

This invention also is directed to methods for producing a bentgrassplant by crossing a first parent bentgrass plant with a second parentbentgrass plant wherein the first or second parent bentgrass plant is abentgrass plant of the variety ‘T-1’. Further, both first and secondparent bentgrass plants can come from the bentgrass variety ‘T-1’. Thus,any such methods using the bentgrass variety ‘T-1’ are part of thisinvention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using bentgrass variety‘T-1’ as a parent are within the scope of this invention, includingthose developed from varieties derived from bentgrass variety ‘T-1’.Advantageously, the bentgrass variety could be used in crosses withother, different, bentgrass plants to produce the first generation (F₁)bentgrass hybrid seeds and plants with superior characteristics. Thevariety of the invention can also be used for transformation whereexogenous genes are introduced and expressed by the variety of theinvention. Genetic variants created either through traditional breedingmethods using variety ‘T-1’ or through transformation of ‘T-1’ by any ofa number of protocols known to those of skill in the art are intended tobe within the scope of this invention.

As used herein, the term plant includes plant cells, plant protoplasts,plant cell tissue cultures from which bentgrass plants can beregenerated, plant calli, plant clumps, and plant cells that are intactin plants or parts of plants, such as embryos, pollen, ovules, flowers,pods, leaves, roots, root tips, anthers, and the like.

Tables

In Table 2 that follows, the traits and characteristics of bentgrasscultivar ‘T-1’ are compared to several competing varieties of commercialbentgrasss of similar maturity. In the table, column 1 shows thevariety; column 2 is the culm length (cm); column 3 gives the flagleaflength (cm); column 4 shows the flagleaf width (mm); column 5 shows theflagleaf length to width ratio (mm); column 6 gives the ligule length(mm); column 7 gives the panicle length (cm); columns 8 and 9 are theseed length and width (mm); column 10 gives the seed length and widthratio (mm); columns 11 and 12 show the seed shape rating and seed colorrating; column 13 gives the seed surface rating; and columns 14 and 15show the seed hairiness rating and awn lemma rating.

In Table 3 below, three different stages of maturity are shown forseveral competing varieties of commercial bentgrass of similar maturity.In the table, column 1 shows the variety; column 2 gives the percent ofplants headed on Jun. 11, 2002; column 3 gives the percent of plants inpollination on Jun. 27, 2002 and column 4 shows the percent of plantswith purple heads on Jun. 27, 2002. TABLE 2 HEAD TO HEAD TRAITCOMPARISON Culm Flagleaf Flagleaf Flagleaf Ligule Panicle Seed Seed Seedlength length width L/W ratio length length length width L/W ratio cm cmmm mm mm cm mm mm mm Alpha 28.3 2.82 2.14 13.9 1.69 5.29 1.53 0.43 3.57Cobra 39.7 4.40 2.83 16.1 2.70 8.17 1.56 0.42 3.80 L-93 40.7 4.41 2.6816.6 1.97 7.88 1.59 0.43 3.71 Penncross 36.6 4.05 2.43 17.6 2.45 7.291.50 0.41 3.67 Pennlinks 32.0 3.61 2.34 16.1 1.84 6.90 1.58 0.42 3.81Providence 39.8 3.95 2.57 16.0 2.10 7.85 1.57 0.44 3.62 Seaside 48.25.95 2.96 20.4 2.77 9.53 1.63 0.43 3.83 Southshore 39.6 3.82 2.58 15.62.63 7.30 1.57 0.43 3.70 SR-1020 37.9 3.40 2.21 16.1 2.29 7.00 1.54 0.433.65 T-1 34.7 3.50 2.47 14.4 1.94 6.67 1.58 0.44 3.62

TABLE 3 HEAD TO HEAD MATURITY COMPARISON Plants headed Plants inpollination Plants with purple Variety Jun. 11, 2002 Jun. 27, 2002 headsJun. 27, 2002 Alpha 77% 88% 44% Cobra 83% 93% 19% L-93 66% 89% 14%Penncross 80% 86% 20% Pennlinks 75% 87% 32% Providence 54% 82% 16%Seaside 21% 58%  8% Southshore 68% 83% 17% SR-1020 46% 80% 17% T-1 93%94% 42%

Deposit Information

A deposit of the bentgrass seed of this invention is maintained by J. R.Simplot Co., 999 Main Street, Boise, Id. 83702. Access to this depositwill be available during the pendency of this application to personsdetermined by the Commissioner of Patents and Trademarks to be entitledthereto under 37 CFR 1.14 and 35 USC 122. Upon allowance of any claimsin this application, all restrictions on the availability to the publicof the variety will be irrevocably removed by affording access to adeposit of at least 2,500 seeds of the same variety with the AmericanType Culture Collection, Manassas, Va. or National Collections ofIndustrial, Food and Marine Bacteria (NCIMB), 23 St Machar Drive,Aberdeen, Scotland, AB24 3RY, United Kingdom.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding. However, it will be obvious that certain changes andmodifications such as single gene modifications and mutations,somoclonal variants, variant individuals selected from large populationsof the plants of the instant variety and the like may be practicedwithin the scope of the invention as limited only by the scope of theappended claims.

1. Seed of bentgrass line designated ‘T-1’, a representative sample ofseed of the line having been deposited under ATCC Accession No. PTA-______.
 2. A bentgrass plant, or a part thereof, produced by growing theseed of claim
 1. 3. A tissue culture of regenerable cells produced fromthe plant of claim
 2. 4. Protoplasts produced from the tissue culture ofclaim
 3. 5. The tissue culture of claim 3, wherein the cells areproduced from a tissue selected from the group consisting of leaves,pollen, embryos, cotyledons, hypocotyls, meristematic cells, roots, roottips, anthers, flowers, stems, culms, stolons and crown tissue.
 6. Abentgrass plant regenerated from the tissue culture of claim 3, whereinthe plant has all the morphological and physiological characteristics ofline ‘T-1’.
 7. A method for producing an F1 hybrid bentgrass seed,comprising crossing the plant of claim 2 with a different bentgrassplant and harvesting the resultant F1 hybrid bentgrass seed.
 8. A hybridbentgrass seed produced by the method of claim
 7. 9. A hybrid bentgrassplant, or a part thereof, produced by growing said hybrid seed of claim8.
 10. A method of producing a bentgrass seed wherein the methodcomprises growing said hybrid bentgrass plant of claim 9 and harvestingthe resultant seed.
 11. A method for producing a male sterile bentgrassplant wherein the method comprises transforming the bentgrass plant ofclaim 2 with a nucleic acid molecule that confers male sterility.
 12. Amale sterile bentgrass plant produced by the method of claim
 11. 13. Amethod of producing an herbicide resistant bentgrass plant wherein themethod comprises transforming the bentgrass plant of claim 2 with atransgene that confers herbicide resistance.
 14. An herbicide resistantbentgrass plant produced by the method of claim
 13. 15. The bentgrassplant of claim 14, wherein the transgene confers resistance to anherbicide selected from the group consisting of: imidazolinone,sulfonylurea, glyphosate, glufosinate, L-phosphinothricin, triazine andbenzonitrile.
 16. A method of producing an insect resistant bentgrassplant wherein the method comprises transforming the bentgrass plant ofclaim 2 with a transgene that confers insect resistance.
 17. An insectresistant bentgrass plant produced by the method of claim
 16. 18. Thebentgrass plant of claim 17, wherein the transgene encodes a Bacillusthuringiensis endotoxin.
 19. A method of producing a disease resistantbentgrass plant wherein the method comprises transforming the bentgrassplant of claim 2 with a transgene that confers disease resistance.
 20. Adisease resistant bentgrass plant produced by the method of claim 19.21. A method of producing a bentgrass plant with modified fatty acidmetabolism or modified carbohydrate metabolism wherein the methodcomprises transforming the bentgrass plant of claim 2 with a transgeneencoding a protein selected from the group consisting of stearyl-ACPdesaturase, fructosyltransferase, levansucrase, alpha-amylase, invertaseand starch branching enzyme.
 22. A bentgrass plant produced by themethod of claim
 21. 23. A bentgrass plant, or part thereof, having allthe physiological and morphological characteristics of the line ‘T-1’, arepresentative sample of seed of the line having been deposited underATCC Accession No. PTA- ______.
 24. A method of introducing a desiredtrait into bentgrass line ‘T-1’ wherein the method comprises: (a)crossing a ‘T-1’ plant, representative seed having been deposited underATCC Accession No. PTA- ______, with a plant of another bentgrass linethat comprises a desired trait to produce F1 progeny plants, wherein thedesired trait is selected from the group consisting of male sterility,herbicide resistance, insect resistance, and resistance to bacterialdisease, fungal disease or viral disease; (b) selecting progeny plantsthat have the desired trait to produce selected progeny plants; (c)crossing the selected progeny plants with the ‘T-1’ plants to producebackcross progeny plants; (d) selecting for backcross progeny plantsthat have the desired trait and physiological and morphologicalcharacteristics of bentgrass line ‘T-1’ listed in Table 1 to produceselected backcross progeny plants; and (e) repeating steps (c) and (d)one or more times in succession to produce selected second or higherbackcross progeny plants that comprise the desired trait and all of thephysiological and morphological characteristics of bentgrass line ‘T-1’listed in Table 1 as determined at the 5% significance level when grownin the same environmental conditions.
 25. A plant produced by the methodof claim 24, wherein the plant has the desired trait and all of thephysiological and morphological characteristics of bentgrass line ‘T-1’listed in Table 1 as determined at the 5% significance level when grownin the same environmental conditions.
 26. The plant of claim 25 whereinthe desired trait is herbicide resistance and the resistance isconferred to an herbicide selected from the group consisting of:imidazolinone, sulfonylurea, glyphosate, glufosinate,L-phosphinothricin, triazine and benzonitrile.
 27. The plant of claim 25wherein the desired trait is insect resistance and the insect resistanceis conferred by a transgene encoding a Bacillus thuringiensis endotoxin.28. The plant of claim 25 wherein the desired trait is male sterilityand the trait is conferred by a cytoplasmic nucleic acid molecule thatconfers male sterility.
 29. A method of modifying fatty acid metabolismor modified carbohydrate metabolism into bentgrass line ‘T-1’ whereinthe method comprises: (a) crossing a ‘T-1’ plant, representative seedhaving been deposited under ATCC Accession No. PTA- ______, with a plantof another bentgrass line to produce F1 progeny plants that comprise anucleic acid molecule encoding an enzyme selected from the groupconsisting of phytase, stearyl-ACP desaturase, fructosyltransferase,levansucrase, alpha-amylase, invertase and starch branching enzyme; (b)selecting progeny plants that have said nucleic acid molecule to produceselected progeny plants; (c) crossing the selected progeny plants withthe ‘T-1’ plants to produce backcross progeny plants; (d) selecting forbackcross progeny plants that have said nucleic acid molecule andphysiological and morphological characteristics of bentgrass line ‘T-1’listed in Table 1 to produce selected backcross progeny plants; and (e)repeating steps (c) and (d) one or more times in succession to produceselected second or higher backcross progeny plants that comprise saidnucleic acid molecule and have all of the physiological andmorphological characteristics of bentgrass line ‘T-1’ listed in Table 1and as determined at the 5% significance level when grown in the sameenvironmental conditions.
 30. A plant produced by the method of claim29, wherein the plant comprises the nucleic acid molecule and has all ofthe physiological and morphological characteristics of bentgrass line‘T-1’ listed in Table 1 and as determined at the 5% significance levelwhen grown in the same environmental conditions.